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Toxoplasma gondii and Eimeria spp.are closely related apicomplexan parasites.T.gondii profilin is the ligand of toll like receptor 4(TLR 4)and activate the pathway of interleukin-12(IL-12)generation in murine DCs.Eimeria maxima is extreme immunogenicity in the seven avian Eimeria spp.parasites and stimulated solid immunity against the parasite re-infection after single oral immunization of its live ooeysts whereas the solid immunity provide by Eimeria tenella need at least two times immunization.However,the mechanism of different immunogenicity in the Eimaeria spp.parasites is not clear.In this study,we constructed a transgenic E.tenella line(Et/EmPro)expressing E.maxima profilin(EmPro)to investigate the immunogenicity of the transgenic parasites.We transfected E.tenella sporozoites with single cassette vector(pSDE-EmProS)based on restriction enzyme-mediate transfection techniques and inoculated the chickens with transgenic sporozoites via cloaca route.We got a transgenic population which about 1.2%sporulated oocysts expressing report gene(enhanced yellow fluorescence protein,EYFP)in the first generation under the selection with pyrimethamine.In the next generations,we selected the transgenic populations based on fluoresce activated cell sorting(FACS)combine with pyrimethamine and more than ninety percent of sporulated oocysts expressing EYFP in the fifth generation.We obtained an approximate 651 bp product after the amplification form the genomic DNA of the fifth generation Et/EmPro with EmPro specific primers,and a 24 kDa sized band after the reaction of Et/EmPro oocysts antigens with anti-flag primary antibody and HRP-coated goat anti-mouse IgG secondary antibody.In addition,we observed EmPro was located in the surface of transgenic sporozoites.Taken together,these results indicated that we got a stable transfected E.tenella population expressing E.maxima profilin for further investigation of the immunogenicity of the transgenic parasites.